Applications of off-gas mass spectrometry in fed-batch mammalian cell culture

Goh, Hai–Yuan; Sulu, Michael; Alosert, Haneen; Lewis, Graham; Josland, G.D.; Merriman, Daniel

doi:10.1007/s00449-019-02242-2

Cited by 12 publications

(14 citation statements)

References 14 publications

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“…The respiratory quotient (RQ), defined as carbon dioxide evolution rate (CER)/oxygen uptake rate (OUR), was also estimated and a distinct correlation between RQ and the metabolic state of the cell culture was established; when a cell culture was determined to be in the lactate production phase, the average RQ was above 1. Conversely, when the culture was determined to be in a lactate consumption phase, the average RQ was below 1 [94,95].…”

Section: Mass Spectrometry Techniquesmentioning

confidence: 95%

“…Correlation between viable cell concentration and oxygen concentration of the inlet gas into the bioreactor was high, irrespective of scale for a CHO-GS cell line that expressed chimeric IgG4 monoclonal antibodies [94]. Additionally, the oxygen mass transfer coefficient (k L a) could also be identified throughout the culture with the mass spectrometer, and an impact of antifoam on k L a was found [94]. The respiratory quotient (RQ), defined as carbon dioxide evolution rate (CER)/oxygen uptake rate (OUR), was also estimated and a distinct correlation between RQ and the metabolic state of the cell culture was established; when a cell culture was determined to be in the lactate production phase, the average RQ was above 1.…”

Section: Mass Spectrometry Techniquesmentioning

confidence: 98%

“…Interestingly, mass spectrometry gas analysis has been realized for fed-batch cultivation of mammalian cell cultures in 5-L and 50-L vessels to measure its application in scale-up systems. Correlation between viable cell concentration and oxygen concentration of the inlet gas into the bioreactor was high, irrespective of scale for a CHO-GS cell line that expressed chimeric IgG4 monoclonal antibodies [94]. Additionally, the oxygen mass transfer coefficient (k L a) could also be identified throughout the culture with the mass spectrometer, and an impact of antifoam on k L a was found [94].…”

Section: Mass Spectrometry Techniquesmentioning

confidence: 98%

See 2 more Smart Citations

Modern Sensor Tools and Techniques for Monitoring, Controlling, and Improving Cell Culture Processes

et al. 2022

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The growing biopharmaceutical industry has reached a level of maturity that allows for the monitoring of numerous key variables for both process characterization and outcome predictions. Sensors were historically used in order to maintain an optimal environment within the reactor to optimize process performance. However, technological innovation has pushed towards on-line in situ continuous monitoring of quality attributes that could previously only be estimated off-line. These new sensing technologies when coupled with software models have shown promise for unique fingerprinting, smart process control, outcome improvement, and prediction. All this can be done without requiring invasive sampling or intervention on the system. In this paper, the state-of-the-art sensing technologies and their applications in the context of cell culture monitoring are reviewed with emphasis on the coming push towards industry 4.0 and smart manufacturing within the biopharmaceutical sector. Additionally, perspectives as to how this can be leveraged to improve both understanding and outcomes of cell culture processes are discussed.

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Section: Mass Spectrometry Techniquesmentioning

confidence: 95%

Section: Mass Spectrometry Techniquesmentioning

confidence: 98%

Section: Mass Spectrometry Techniquesmentioning

confidence: 98%

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Modern Sensor Tools and Techniques for Monitoring, Controlling, and Improving Cell Culture Processes

et al. 2022

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“…Over the past several years, numerous spectroscopic methods (e.g., Raman, near-infrared [NIR], fluorescence), capacitance sensors, turbidity measurements as well as advanced off-gas analysis have been adopted as PAT in biopharmaceutical manufacturing to monitor and control process performance (Goh et al, 2020;Kroll et al, 2017;Moore et al, 2019;Santos et al, 2018). These methods provide substantial information about multiple process and metabolite variables, while being non-invasive and non-destructive, hence representing excellent candidates for analysis of cell culture processes.…”

Section: Upstreammentioning

confidence: 99%

“…The in-situ combi-sensor was shown to be capable of determining qOUR together with turbidity measurement for the estimation of cell concentration for various hosts (Dahlmann et al, 2020). Another study showed that off-gas analysis using a magnetic sector mass spectrometer delivered reliable values for specific oxygen uptake rates for both bench and pilot scale cultivations (Goh et al, 2020). Furthermore, the authors correlated the respiratory quotient (RQ) to the metabolic state and viable cell density and differed between lactate production (average RQ > 1) and consumption phase (average RQ < 1).…”

Section: Upstreammentioning

confidence: 99%

Systematic assessment of process analytical technologies for biologics

Gillespie

Wasalathanthri

Ritz

et al. 2021

Biotech & Bioengineering

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The application of process analytical technology (PAT) for biotherapeutic development and manufacturing has been employed owing to technological, economic, and regulatory advantages across the industry. Typically, chromatographic, spectroscopic, and/or mass spectrometric sensors are integrated into upstream and downstream unit operations in in-line, on-line, or at-line fashion to enable real-time monitoring and control of the process. Despite the widespread utility of PAT technologies at various unit operations of the bioprocess, a holistic business value assessment of PAT has not been well addressed in biologics. Thus, in this study, we evaluated PAT technologies based on predefined criteria for their technological attributes such as enablement of better process understanding, control, and highthroughput capabilities; as well as for business attributes such as simplicity of implementation, lead time, and cost reduction. The study involved an industry-wide survey, where input from subject matter industry experts on various PAT tools were collected, assessed, and ranked. The survey results demonstrated on-line liquid Chromatography (LC), in-line Raman, and gas analysis techniques are of high business value especially at the production bioreactor unit operation of upstream processing. In-line variable path-length UV/VIS measurements (VPE), on-line LC, multiangle light scattering (MALS), and automated sampling are of high business value in Protein A purification and polishing steps of the downstream process. We also provide insights, based on our experience in clinical and commercial manufacturing of biologics, into the development and implementation of some of the PAT tools. The results presented in this study are intended to be helpful for the current practitioners of PAT as well as those new to the field to gauge, prioritize and steer their projects for success.

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Non‐invasive and time‐resolved measurement of the respiration activity of Chinese hamster ovary cells enables prediction of key culture parameters in shake flasks

Ihling

Munkler

Paul

et al. 2022

Biotechnology Journal

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Background: Shake flasks are frequently used for mammalian cell suspension cultures. For process development and routine culture monitoring, information on culture behavior is needed early on. Main methods and major results:Here, cell-specific oxygen uptake rates (qO 2 ) of two CHO cell lines were determined from shake flask experiments by simultaneous measurement of oxygen transfer rates (OTR) and viable cell concentrations (VCC). For cell line one, qO 2 decreased from 2.38⋅10 −10 to 1.02⋅10 -10 mmol cell −1 h −1 during batch growth. For cell line two, qO 2 was constant (1.90⋅10 −10 mmol h −1 ). Determined qO 2 values were used to calculate the VCC from OTR data. Cumulated oxygen consumption and glucose consumption were correlated for both cell lines and enabled calculation of glucose concentrations from OTR data. IgG producing cell line one had an oxygen demand of ∼15 mmol oxygen g glucose −1 , cell line two consumed ∼5 mmol oxygen g glucose −1 .The established correlations for determination of VCC and glucose were successfully transferred to subsequent cultivations for both cell lines. Combined measurement of the OTR and the carbon dioxide transfer rate enabled quantitative determination of the lactate concentration (production and consumption) without sampling. Conclusions and implications:Taken together, non-invasive measurement of the respiration activity enabled time-resolved determination of key culture parameters for increased process understanding in shake flasks. K E Y W O R D Scarbon dioxide transfer rate, Chinese hamster ovary cells, oxygen transfer rate, respiratory quotient, shake flasks INTRODUCTIONChinese hamster ovary (CHO) cells are the most important mammalian cells used for the stable production of therapeutic proteins. [1] Even though traditionally applied for microbial cultivations, shake flasks areThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Applications of off-gas mass spectrometry in fed-batch mammalian cell culture

Cited by 12 publications

References 14 publications

Modern Sensor Tools and Techniques for Monitoring, Controlling, and Improving Cell Culture Processes

Modern Sensor Tools and Techniques for Monitoring, Controlling, and Improving Cell Culture Processes

Systematic assessment of process analytical technologies for biologics

Non‐invasive and time‐resolved measurement of the respiration activity of Chinese hamster ovary cells enables prediction of key culture parameters in shake flasks

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